US4910139A - Method for continuously producing citric acid by dual hollow fiber membrane bioreactor - Google Patents

Method for continuously producing citric acid by dual hollow fiber membrane bioreactor Download PDF

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US4910139A
US4910139A US07/226,718 US22671888A US4910139A US 4910139 A US4910139 A US 4910139A US 22671888 A US22671888 A US 22671888A US 4910139 A US4910139 A US 4910139A
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citric acid
hollow fiber
bioreactor
fiber membrane
culture medium
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Ho Nam Chang
Bong Hyun Chung
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Korea Advanced Institute of Science and Technology KAIST
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/10—Hollow fibers or tubes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/16—Hollow fibers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids
    • C12P7/48—Tricarboxylic acids, e.g. citric acid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/66—Aspergillus
    • C12R2001/685—Aspergillus niger
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/8215—Microorganisms
    • Y10S435/911—Microorganisms using fungi
    • Y10S435/913—Aspergillus
    • Y10S435/917—Aspergillus niger

Definitions

  • the present invention relates to the method for producing citric acid on a continuous basis, after fixing Aspergillus niger B60 as mildews, in a dual hollow fiber membrane bioreactor.
  • citric acid is commonly produced by fermentation, 20 percent of them is produced by surface culture and remaining 80 percent by submerged culture (Sodeck et al., Proc. Biochem. Oct./Nov., 9, 1981).
  • methods according to the prior art are batch operation which have a disadvantage of lower productivity in comparison with continuous operation according to present invention.
  • As the existing continuous methods improved on the batch method there are (1) a method utilizing a fermenter equipped with an agitator (Kristiansen and Sinclair, Biotechnol. Bioeng., 21, 297, 1979), (2) a method utilizing air-lift reactor after fixing Aspergillus niger mycelium to alginate bead (Vaija et al., Appl. Biochem.
  • the method according to the present invention has overcome the above described difficulties by utilizing dual hollowfiber membrane bioreactor, and obtained higher citric acid concentrations than the batch operation as well as higher productivity.
  • cells are separated from the product by a membrane between them so that eliminate primary purification process which is essential in the ordinary fermentation process. Therefore, the present invention has processwise advantage saving considerable amount of energy required for aeration and agitation in the case of ordinary fermentation.
  • a dual hollow fiber membrane type biological bioreactor had been made by modifying the existing hollowfiber membrane bioreactor for the culture of aerobic bacteria.
  • Robertson and Kim inserted three silicone tubes into a polypropylene hollowfiber membrane to supply liquid nutrients outside the polypropylene membrane, while oxygen into silicon tubes. Culturing Aerobic Streptomyces aureofaciens bacteria between them they carried out the researches related to continuous production of tetracycline (Robertson and Kim, Biotechnol. Bioeng., 27, 1012, 1985).
  • the inventors of the present invention have achieved successfully continuous production of rifamycin B at a first time by utilizing the bioreactor, unlike the bioreactor used by Robertson and Kim, which comprises a silicone tube for supplying oxygen to its outside, and three polypropylene hollow fiber membranes inserted into the silicone tube for feeding liquid nutrients, and Nocardia mediterranei is cultured between the tubes (Chan et al., ACS Symp. Ser. 314, 32, 1986).
  • the present invention is related to a method of producing citric acid on a continuous basis by utilizing the bioreactor developed by the present inventors and the details of the method will be described hereinafter with reference to the accompanying drawings.
  • FIG. 1 is a sectional view of a dual hollow fiber membrane bioreactor
  • FIG. 2 is a transverse sectional view of a double fine-tube reactor used in the present invention.
  • FIG. 3 is a three-dimensional microphotograph of the cross section of a dual hollow fiber membrane bioreactor and after the culture of cells.
  • FIG. 1 is a sectional view of a dual hollowfiber membrane of the bioreactor according to the present invention.
  • three polypropylene hollowfiber membranes (2) are inside a silicone tube (1), while Aspergillus niger B60 (5) is contained in the space between the silicon tube (1) and three polypropylene hollowfiber membrane (2).
  • FIG. 2 is a transverse sectional view of the dual hollow fiber membrane bioreactor (10) which is used in the present invention.
  • the dual hollow fiber membranes of FIG. 1, where polypropylene tubes (2) are longer than the silicone tube (1) are laid in a glass tube (6) in parallel and then the ends of the silicone tube (1) and polypropylene fine tubes (2) are fixed with silicone rubber (7) respectively.
  • an oxygen supply inlet (3a) is provided at one end of the silicone tube (1) while an air vent (3b) is provided at the other end.
  • a three-way valve (9) is provided between one end of the silicone tube (1) and polypropylene fine tubes (2) and a culture strain inlet (8) on the other side.
  • One end of the glass tube is fitted with a liquid culture medium inlet (4a) while a culture medium and product citric acid outlet (4b) is fitted on the other side.
  • liquid culture medium (4) is fed in the direction of a liquid culture medium and citric acid discharge outlet (4b) through a liquid culture medium inlet (4a), while oxygen (3) is supplied in the direction of an air vent (3b) from an oxygen feed inlet (3a).
  • the concentration of cells in the bioreactor (10) is equalized by alternately changing the feeding direction of liquid culture medium by means of a three-way valve (9). After the growth of cells comes to an end, the liquid culture medium deficient in nitrogen source is supplied to ensure stable operation of the bioreactor and also higher productivity.
  • the polypropylene fine tubes used in the fabrication of the bioreactor in the present invention where Enka items of Germany, measuring 0.033 cm in inner diameter and 0.063 cm in outer diameter, with pore sizes ranging from 0.4 to 0.6 ⁇ m.
  • the silicone tube was Dow Corning product of the United States, with its inner diameter is 0.147 cm and outer diameter 0.196 cm. Eight double fine tubes were laid in a glass tube of 0.8 cm inner diameter, and the length of their possible contact with oxygen was 16 cm.
  • the scope of the present invention is not limited to the bioreactor used herein. That is, materials other than silicone and polypropylene can be used in the fabrication of a bioreactor and its structure can be also diverse depending upon the specification of fine tubes to be used, the number of fine tubes to be inserted can be widely adjusted.
  • FIG. 3 is a three-dimensional microphotograph showing the cross section of a dual hollow fiber membrane before (left) and after (right) the culture of cells. As shown in the microphotograph, Aspergillus niger B60 is cultured in high concentration in the space between the silicone tube (1) and polypropylene hollow fiber membrane (2) by feeding oxygen and liquid culture medium.
  • Aspergillus niger B60 was inoculated to a 500-ml triangular flask containing 100 ml of liquid culture medium (Composition: sugar 6 g, NH 4 NO 3 0.25 g, KH 2 PO 4 0.1 g, MgSO 4 .7H 2 O 0.025 g, pH 3.1) to be adjusted to an initial spore concentration of 10 7 -10 8 spores per 100 ml and then it was cultured in a shaking incubator at 300 rpm and 30° C.
  • liquid culture medium Composition: sugar 6 g, NH 4 NO 3 0.25 g, KH 2 PO 4 0.1 g, MgSO 4 .7H 2 O 0.025 g, pH 3.1
  • citric acid was measured by the method of Marier and Boulet (Marier and Boulet, J. Dairy Sci., 41, 1683, 1958), while sugar concentration was determined by the method of Dobois and others for the total sugar concentration in culture medium (Dubois et al., Anal. Chem. 28, 350, 1956) on the basis of cane sugar and found that 18 g/liter of citric acid was produced, with its yield and productivity standing at 40 percent and 0.06 g/liter hour, respectively.
  • the polypropylene fine tubes (2) of FIG. 2 were dipped in 50% ethanol and sterilized with 5% formalin and then sufficiently washed with sterilized distilled water, after then, Aspergillus niger B60 cultured in a shaking incubator for seven days was inoculated in the space between the silicone tube (1) and polypropylene fine tubes (2) through a culture strain inlet (8) as shown in FIG. 2. It was supplied with oxygen in the direction of an air vent (3b) from an oxygen inlet (3a) at 28°-30° C. for 10 days, while the liquid culture medium as used in the comparative example was fed in the direction of a citric acid discharge outlet (4b) from a liquid culture medium inlet (4a), to cultivate the cells in high concentration. The feeding direction of liquid culture medium was alternately changed by means of three way valve to equalize the concentration of cells in the bioreactor.
  • the product was collected from a citric acid outlet (4b), while continuously feeding the culture medium deficient in NH 4 NO 3 and the collected citric acid was measured.
  • the product was 6.5 g/liter at the flow rate of 2 ml/hour. Its productivity was 1.62 g/liter hour, and 27 times of the comparative example.

Abstract

Aspergillus niger B60 was immobilized in a dual hollow fiber bioreactor to produce citric acid continuously. The fungi proliferated well in the interstitial region formed by a parallel arrangement of three microporous polypropylene hollow fibers contained within a silicone tube. Excessive fungal cell growth could be controlled by supplying a nitrogen-deficient medium at the production stage, and also uniform cell distribution in the reactor could be obtained by changing the direction of medium flow. With pure oxygen aeration and nitrogen-deficient medium, volumetric productivity reached 1.62 g/l.h at a residence time of 4.02 h, which corresponded to a 27-fold increase over that of shake-flask fermentation. When the residence time was increased to 20.1 h, citric acid at a concentration of 26 g/l was continuously produced, with a yield of 80-90% and a volumetric productivity of 1.3 g/l.h. This represents a significant improvement in final concentration, yield and the volumetric productivity over the equivalent values of the corresponding batch formentation, which were 18 g/l, 40% and 0.06 g/l.h, respectively.

Description

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to the method for producing citric acid on a continuous basis, after fixing Aspergillus niger B60 as mildews, in a dual hollow fiber membrane bioreactor.
2. Description of the Related Art
Recently, citric acid is commonly produced by fermentation, 20 percent of them is produced by surface culture and remaining 80 percent by submerged culture (Sodeck et al., Proc. Biochem. Oct./Nov., 9, 1981). However, such methods according to the prior art are batch operation which have a disadvantage of lower productivity in comparison with continuous operation according to present invention. As the existing continuous methods improved on the batch method, there are (1) a method utilizing a fermenter equipped with an agitator (Kristiansen and Sinclair, Biotechnol. Bioeng., 21, 297, 1979), (2) a method utilizing air-lift reactor after fixing Aspergillus niger mycelium to alginate bead (Vaija et al., Appl. Biochem. Biotechnol., 7, 51, 1982) and (3) a method utilizing tower reactor after the fixation of Aspergillus niger to polyacrylamide (Horitsu et al., Appl. Microbiol. Biotechnol., 22, 8, 1985).
The continuous operations in the existing method (1)˜(3) brought higher productivity, but less concentration than the batch operation as in the case of other continuous operations, and required higher costs for the process of purification.
Moreover, when carriers are used to fix mildews, there was a tendency to weaken the carriers due to an excessive growth of cells. Leaving the carriers, cells, also, grow in liquid culture medium to cause many operational difficulties in the case of continuous operation.
SUMMARY OF THE INVENTION
The method according to the present invention has overcome the above described difficulties by utilizing dual hollowfiber membrane bioreactor, and obtained higher citric acid concentrations than the batch operation as well as higher productivity. In accordance with the present invention, cells are separated from the product by a membrane between them so that eliminate primary purification process which is essential in the ordinary fermentation process. Therefore, the present invention has processwise advantage saving considerable amount of energy required for aeration and agitation in the case of ordinary fermentation.
A dual hollow fiber membrane type biological bioreactor had been made by modifying the existing hollowfiber membrane bioreactor for the culture of aerobic bacteria. Robertson and Kim inserted three silicone tubes into a polypropylene hollowfiber membrane to supply liquid nutrients outside the polypropylene membrane, while oxygen into silicon tubes. Culturing Aerobic Streptomyces aureofaciens bacteria between them they carried out the researches related to continuous production of tetracycline (Robertson and Kim, Biotechnol. Bioeng., 27, 1012, 1985).
After then, the inventors of the present invention have achieved successfully continuous production of rifamycin B at a first time by utilizing the bioreactor, unlike the bioreactor used by Robertson and Kim, which comprises a silicone tube for supplying oxygen to its outside, and three polypropylene hollow fiber membranes inserted into the silicone tube for feeding liquid nutrients, and Nocardia mediterranei is cultured between the tubes (Chan et al., ACS Symp. Ser. 314, 32, 1986).
The present invention is related to a method of producing citric acid on a continuous basis by utilizing the bioreactor developed by the present inventors and the details of the method will be described hereinafter with reference to the accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a sectional view of a dual hollow fiber membrane bioreactor;
FIG. 2 is a transverse sectional view of a double fine-tube reactor used in the present invention; and
FIG. 3 is a three-dimensional microphotograph of the cross section of a dual hollow fiber membrane bioreactor and after the culture of cells.
DESCRIPTION OF THE PREFERRED EMBODIMENTS
FIG. 1 is a sectional view of a dual hollowfiber membrane of the bioreactor according to the present invention. In the drawing, three polypropylene hollowfiber membranes (2) are inside a silicone tube (1), while Aspergillus niger B60 (5) is contained in the space between the silicon tube (1) and three polypropylene hollowfiber membrane (2).
FIG. 2 is a transverse sectional view of the dual hollow fiber membrane bioreactor (10) which is used in the present invention. As shown in the drawing, the dual hollow fiber membranes of FIG. 1, where polypropylene tubes (2) are longer than the silicone tube (1), are laid in a glass tube (6) in parallel and then the ends of the silicone tube (1) and polypropylene fine tubes (2) are fixed with silicone rubber (7) respectively. At one end of the silicone tube (1), an oxygen supply inlet (3a) is provided while an air vent (3b) is provided at the other end. Between one end of the silicone tube (1) and polypropylene fine tubes (2) a three-way valve (9) is provided and a culture strain inlet (8) on the other side. One end of the glass tube is fitted with a liquid culture medium inlet (4a) while a culture medium and product citric acid outlet (4b) is fitted on the other side.
After inoculating bacteria in the space between the inside of the silicone tube (1) and the outside of polypropylene tubes (2) through a culture strain inlet (8), liquid culture medium (4) is fed in the direction of a liquid culture medium and citric acid discharge outlet (4b) through a liquid culture medium inlet (4a), while oxygen (3) is supplied in the direction of an air vent (3b) from an oxygen feed inlet (3a).
The concentration of cells in the bioreactor (10) is equalized by alternately changing the feeding direction of liquid culture medium by means of a three-way valve (9). After the growth of cells comes to an end, the liquid culture medium deficient in nitrogen source is supplied to ensure stable operation of the bioreactor and also higher productivity.
The polypropylene fine tubes used in the fabrication of the bioreactor in the present invention where Enka items of Germany, measuring 0.033 cm in inner diameter and 0.063 cm in outer diameter, with pore sizes ranging from 0.4 to 0.6 μm. The silicone tube was Dow Corning product of the United States, with its inner diameter is 0.147 cm and outer diameter 0.196 cm. Eight double fine tubes were laid in a glass tube of 0.8 cm inner diameter, and the length of their possible contact with oxygen was 16 cm.
However, the scope of the present invention is not limited to the bioreactor used herein. That is, materials other than silicone and polypropylene can be used in the fabrication of a bioreactor and its structure can be also diverse depending upon the specification of fine tubes to be used, the number of fine tubes to be inserted can be widely adjusted.
FIG. 3 is a three-dimensional microphotograph showing the cross section of a dual hollow fiber membrane before (left) and after (right) the culture of cells. As shown in the microphotograph, Aspergillus niger B60 is cultured in high concentration in the space between the silicone tube (1) and polypropylene hollow fiber membrane (2) by feeding oxygen and liquid culture medium.
In accordance with the present invention, it is possible to obtain excellent results which are higher by 40-50 percent in yield and 22-27 times in productivity than those in the batch operation in a conventional shaking incubator. It has an advantage of easy separation and purification since citric acid is produced continuously by fine tubes under separate state from cells.
Following examples are descriptions of the conventional method (comparative embodiment) and the present invention. But the present invention is not limited to these examples.
Comparative Example
Aspergillus niger B60 was inoculated to a 500-ml triangular flask containing 100 ml of liquid culture medium (Composition: sugar 6 g, NH4 NO3 0.25 g, KH2 PO4 0.1 g, MgSO4.7H2 O 0.025 g, pH 3.1) to be adjusted to an initial spore concentration of 107 -108 spores per 100 ml and then it was cultured in a shaking incubator at 300 rpm and 30° C.
After 10 days of incubation, the volume of citric acid was measured by the method of Marier and Boulet (Marier and Boulet, J. Dairy Sci., 41, 1683, 1958), while sugar concentration was determined by the method of Dobois and others for the total sugar concentration in culture medium (Dubois et al., Anal. Chem. 28, 350, 1956) on the basis of cane sugar and found that 18 g/liter of citric acid was produced, with its yield and productivity standing at 40 percent and 0.06 g/liter hour, respectively.
Example
The polypropylene fine tubes (2) of FIG. 2 were dipped in 50% ethanol and sterilized with 5% formalin and then sufficiently washed with sterilized distilled water, after then, Aspergillus niger B60 cultured in a shaking incubator for seven days was inoculated in the space between the silicone tube (1) and polypropylene fine tubes (2) through a culture strain inlet (8) as shown in FIG. 2. It was supplied with oxygen in the direction of an air vent (3b) from an oxygen inlet (3a) at 28°-30° C. for 10 days, while the liquid culture medium as used in the comparative example was fed in the direction of a citric acid discharge outlet (4b) from a liquid culture medium inlet (4a), to cultivate the cells in high concentration. The feeding direction of liquid culture medium was alternately changed by means of three way valve to equalize the concentration of cells in the bioreactor.
Further, the product was collected from a citric acid outlet (4b), while continuously feeding the culture medium deficient in NH4 NO3 and the collected citric acid was measured. The product was 6.5 g/liter at the flow rate of 2 ml/hour. Its productivity was 1.62 g/liter hour, and 27 times of the comparative example.
After 15 days, the flow rate was changed to 0.9 ml/h, and the product was 13.5 g/liter. Its yield and productivity were 80-90 percent and 1.5 g/liter hour respectively. Changing the flow rate to 0.4 ml/h after 24 days, 26 g/liter of citric acid was produced while its yield and productivity registered 80-90 percent and 1.3 g/liter hour, respectively, and 22 times of the comparative example.

Claims (2)

What is claimed is:
1. A method for continuously producing citric acid utilizing a dual hollow fiber membrane bioreactor comprising the steps:
inoculating Aspergillus niger B60 in the space between hollow fibers and outer tube of said bioreactor,
culturing said Aspergillus niger B60 in high concentration by feeding liquid culture medium into said hollow fibers supplying oxygen outside of said outer tube, and then,
feeding said liquid culture medium which contains no NH4 NO3 into said hollow fiber membrane, supplying oxygen outside of said outer tube.
2. A method according to claim 1, the feeding direction of said liquid culture medium being changeable alternately by means of a three way valve to equalize the high concentration of Aspergillus niger B60 to be cultured in the bioreactor.
US07/226,718 1987-09-09 1988-08-01 Method for continuously producing citric acid by dual hollow fiber membrane bioreactor Expired - Lifetime US4910139A (en)

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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5045459A (en) * 1990-10-05 1991-09-03 Haarmann & Reimer Corp. Method for the production of granular citric acid
US5104799A (en) * 1990-10-05 1992-04-14 Haarmann & Reimer Method for the production of granular citric acid and salts thereof
US5149643A (en) * 1990-10-05 1992-09-22 Haarmann & Beimer Method for the production of granular citric acid and salts thereof
US5164081A (en) * 1989-03-24 1992-11-17 The Standard Oil Company Apparatus for separation and for treatment of fluid feedstreams, wafers for use therein and related methods
US5174900A (en) * 1989-03-24 1992-12-29 The Standard Oil Company Apparatus for separation and for treatment of fluid feedstreams, wafers for use therein and related methods
US20020146430A1 (en) * 2000-11-22 2002-10-10 Galen James E. Use of ClyA hemolysin for excretion of proteins
US20060293261A1 (en) * 2005-06-24 2006-12-28 Taisho Pharmaceutical Co., Ltd. Clarithromycin or a salt thereof for the treatment or prevention of pulmonary disorders caused by the destruction of pulmonary alveoli
US20070122904A1 (en) * 2000-09-29 2007-05-31 Unisearch Limited Method and apparatus for culturing cells
WO2009149083A2 (en) 2008-06-03 2009-12-10 University Of Maryland, Baltimore Non-hemolytic clya for excretion of proteins
US20100159555A1 (en) * 2005-06-30 2010-06-24 Synexa Life Sciences (Proprietary) Limited Production of secondary metabolites using capillary membranes
US9677042B2 (en) 2010-10-08 2017-06-13 Terumo Bct, Inc. Customizable methods and systems of growing and harvesting cells in a hollow fiber bioreactor system
US11624046B2 (en) 2017-03-31 2023-04-11 Terumo Bct, Inc. Cell expansion
US11629332B2 (en) 2017-03-31 2023-04-18 Terumo Bct, Inc. Cell expansion
US11667881B2 (en) 2014-09-26 2023-06-06 Terumo Bct, Inc. Scheduled feed

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH02255079A (en) * 1989-03-29 1990-10-15 Shimadzu Corp Cell culture apparatus
JPH02276565A (en) * 1989-04-18 1990-11-13 Japanese Res & Dev Assoc Bio Reactor Syst Food Ind Apparatus for continuous enzymic reaction

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU907072A1 (en) * 1980-06-30 1982-02-23 Ленинградский межотраслевой научно-исследовательский институт пищевой промышленности Method for preparing citric acid
FR2507500A1 (en) * 1981-06-12 1982-12-17 Dumont Engineering Et Cie Sa Rotary agitator - aerator partic for prodn. of metabolites etc. - using rotor contg. air compressing piston reciprocated by eccentric

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU907072A1 (en) * 1980-06-30 1982-02-23 Ленинградский межотраслевой научно-исследовательский институт пищевой промышленности Method for preparing citric acid
FR2507500A1 (en) * 1981-06-12 1982-12-17 Dumont Engineering Et Cie Sa Rotary agitator - aerator partic for prodn. of metabolites etc. - using rotor contg. air compressing piston reciprocated by eccentric

Non-Patent Citations (16)

* Cited by examiner, † Cited by third party
Title
Biotech, 85 02469, Mahala et al., World Biotech Rep., 1984, 2, A129 140. *
Biotech, 85-02469, Mahala et al., World Biotech Rep., 1984, 2, A129-140.
Biotech, 87 01837, Chang et al., ACS Symp., (1986), 314. *
Biotech, 87 08034, Eikmeier Z. Naturforsch C, (1987), 42, 4, 408 13. *
Biotech, 87-01837, Chang et al., ACS Symp., (1986), 314.
Biotech, 87-08034, Eikmeier Z. Naturforsch C, (1987), 42, 4, 408-13.
Biotech, 88 0857, Chung et al., Biotech Bioeng, 1988, 32, 2, 205 12. *
Biotech, 88-0857, Chung et al., Biotech Bioeng, 1988, 32, 2, 205-12.
Derwent Abs., 78 71922A/40, Metkin et al., SU 579303, (11 1978). *
Derwent Abs., 78-71922A/40, Metkin et al., SU 579303, (11-1978).
Derwent Abs., 79 45384 B/24, Metkin et al., SU 619511, (7 1978). *
Derwent Abs., 79-45384 B/24, Metkin et al., SU-619511, (7-1978).
Derwent Abs., 81 00326D/01, Blain et al., GB 1581832, (12 1980). *
Derwent Abs., 81-00326D/01, Blain et al., GB 1581832, (12-1980).
Derwent Abstract, 82 64080E/31, Eysmond Fr 2496122, (6 82). *
Derwent Abstract, 82-64080E/31, Eysmond Fr 2496122, (6-82).

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US5045459A (en) * 1990-10-05 1991-09-03 Haarmann & Reimer Corp. Method for the production of granular citric acid
US5104799A (en) * 1990-10-05 1992-04-14 Haarmann & Reimer Method for the production of granular citric acid and salts thereof
US5149643A (en) * 1990-10-05 1992-09-22 Haarmann & Beimer Method for the production of granular citric acid and salts thereof
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US20020146430A1 (en) * 2000-11-22 2002-10-10 Galen James E. Use of ClyA hemolysin for excretion of proteins
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